title: Induction of vesicular steatosis by amiodarone and tetracycline is associated with up-regulation of lipogenic genes in HepaRG cells.
abstract: Drug-induced liver injury occurs in general after several weeks and is often unpredictable. It is characterized by a large spectrum of lesions that includes steatosis and phospholipidosis. Many drugs such as amiodarone and tetracycline have been reported to cause phospholipidosis and/or steatosis. In this study, acute and chronic hepatic effects of these two drugs were investigated using well-differentiated human hepatoma HepaRG cells. Accumulation of typical lipid droplets, labeled with Oil Red O, was observed in hepatocyte-like HepaRG cells after repeat exposure to either drug. Amiodarone caused the formation of additional intracytoplasmic vesicles that did not stain in all HepaRG cells. At the electron microscopic level, these vesicles appeared as typical lamellar bodies and were associated with an increase of phosphatidylethanolamine and phosphatidylcholine. A dose-dependent induction of triglycerides (TG) was observed after repeat exposure to either amiodarone or tetracycline. Several genes known to be related to lipogenesis were induced after treatment by these two drugs. By contrast, opposite deregulation of some of these genes (FASN, SCD1, and THSRP) was observed in fat HepaRG cells induced by oleic acid overload, supporting the conclusion that different mechanisms were involved in the induction of steatosis by drugs and oleic acid. Moreover, several genes related to lipid droplet formation (ADFP, PLIN4) were up-regulated after exposure to both drugs and oleic acid.
  
  
  CONCLUSIONS
  Our results show that amiodarone causes phospholipidosis after short-term treatment and, like tetracycline, induces vesicular steatosis after repeat exposure in HepaRG cells. These data represent the first demonstration that drugs can induce vesicular steatosis in vitro and show a direct relationship between TG accumulation and enhanced expression of lipogenic genes.
authors.affiliation: INSERM UMR 991, Rennes, France.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Anthérieu
authors.forename: Alexandra
authors.initials: A
authors.lastname: Rogue
authors.forename: Bernard
authors.initials: B
authors.lastname: Fromenty
authors.forename: André
authors.initials: A
authors.lastname: Guillouzo
authors.forename: Marie-Anne
authors.initials: MA
authors.lastname: Robin
mesh_terms: D000638:Amiodarone / Q000009:adverse effects / Q000494:pharmacology*; D006528:Carcinoma, Hepatocellular / Q000378:metabolism* / Q000473:pathology; D002352:Carrier Proteins; D045744:Cell Line, Tumor; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D054890:Fatty Acid Synthase, Type I / Q000235:genetics / Q000378:metabolism; D005234:Fatty Liver / Q000139:chemically induced / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D050356:Lipid Metabolism / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D050155:Lipogenesis / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D008113:Liver Neoplasms / Q000378:metabolism* / Q000473:pathology; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D019301:Oleic Acid / Q000494:pharmacology; D000070780:Perilipin-2; D000071229:Perilipin-4; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism; D013230:Stearoyl-CoA Desaturase / Q000235:genetics / Q000378:metabolism; D013752:Tetracycline / Q000009:adverse effects / Q000494:pharmacology*; D014280:Triglycerides / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects* / Q000235:genetics / Q000502:physiology
chemical_list: D002352:Carrier Proteins; D008565:Membrane Proteins; C000602982:PLIN2 protein, human; C560544:PLIN4 protein, human; D000070780:Perilipin-2; D000071229:Perilipin-4; D010750:Phosphoproteins; D014280:Triglycerides; D019301:Oleic Acid; C523776:SCD1 protein, human; D013230:Stearoyl-CoA Desaturase; C518729:FASN protein, human; D054890:Fatty Acid Synthase, Type I; D013752:Tetracycline; D000638:Amiodarone